■本研究旨在研究“谷氨酸(Glu)-γ-氨基丁酸(GABA)”环的病理变化,并应用广泛的靶向代谢组学分析技术来全面探索抽动障碍(TD)大鼠丘脑中的代谢异常。
■将Wistar大鼠随机分为对照组,TD,和tiapride(Tia)组。在大鼠中使用亚氨基二丙腈(IDPN)诱导TD。Tia组给予硫必利。使用UPLC-3QMS测量三组大鼠丘脑中的神经递质水平。And,采用蛋白质印迹法检测Glu脱羧酶(GAD65/67)和GABA转运蛋白(GAD-T)的蛋白表达水平。使用实时聚合酶链反应评估这些基因的mRNA表达水平。最后,通过广泛靶向的代谢组学分析在TD组和对照组大鼠之间检测丘脑中的其他代谢物。
■Glu水平,Glu/GABA比值,和Asp水平在TD组显著高于对照组(均p<0.001),而GABA和Gly水平较低(分别为p<0.001和p=0.009)。与TD组相比,Tia组的Glu水平显着降低(p=0.001)。TD组GAD67蛋白表达水平高于对照组(p=0.009),GAD65、GAD67和GAT-1mRNA表达水平低于对照组(p<0.05)。Tia组的GAD65、GAD67或GAT-1表达没有任何差异。广泛靶向的代谢组学分析显示,TD组和对照组之间有34种物质异常(9种上调,25种下调)。神经类固醇(孕酮,皮质酮)表现出明显的差异。使用京都百科全书对基因和基因组的代谢分析表明,类固醇激素生物合成途径可能与TD发病机理有关。
■这项研究揭示了TD大鼠丘脑的代谢异常。神经递质与神经类固醇生物合成之间的相互作用代表了未来研究的新方向。
UNASSIGNED: This study aimed to investigate pathological changes in the \"Glutamate (Glu)-γ-aminobutyric acid (GABA)\" loop and apply widely targeted metabolomic analysis technology to comprehensively explore metabolite abnormalities/ in the thalamus of rats with tic disorders (TD).
UNASSIGNED: Wistar rats were randomized into control, TD, and tiapride (Tia) groups. Iminodipropionitrile (IDPN) was used to induce TD in rats. The Tia group was administered tiapride. Neurotransmitter levels in the thalamus of rats in the three groups were measured using UPLC-3Q MS. And, the protein expression levels of Glu decarboxylase (GAD65/67) and GABA transporter protein (GAD-T) were measured using western blotting. The mRNA expression levels of these genes were evaluated using real-time polymerase chain reaction. Lastly, other metabolites in the thalamus were detected by widely targeted metabolomic analysis between TD and Control group rats.
UNASSIGNED: The Glu level, Glu/GABA ratio, and Asp level in the TD group were significantly higher (all p < 0.001) than those of the Control group, whereas the GABA and Gly levels were lower (p < 0.001 and p = 0.009, respectively). The Tia group exhibited a significant reduction in the Glu level (p = 0.001) compared with the TD group. The protein expression level of GAD67 in TD group was higher (p = 0.009) and the mRNA expression levels of GAD65, GAD67, and GAT-1 were lower (p < 0.05) than those of the Control group. The Tia group did not display any differences in GAD65, GAD67, or GAT-1 expression. Widely targeted metabolomic analysis revealed that 34 substances were abnornal between the TD and Control groups (9 upregulated and 25 downregulated). Neurosteroids (progesterone, corticosterone) exhibited distinct differences. Metabolite analysis using the Kyoto encyclopedia for genes and genomes indicated that the steroid hormone biosynthesis pathway may be involved in TD pathogenesis.
UNASSIGNED: This study revealed metabolic abnormalities in the thalamus of rats with TD. The interaction between neurotransmitters and neurosteroid biosynthesis represents a new direction for future studies.